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PubMed1992

[Peritonitis caused by Nocardia farcinica in a patient undergoing continuous ambulatory peritoneal dialysis (note)].

We report here an isolation of bacteria genus Nocardia, Nocardia farcinica, in a dialysis liquid of a renal insufficiency patient treated by continuous ambulatory peritoneal dialysis (C.A.P.D.). Real peritonitis was due to infection with this bacteria, eliminated after an adapted antibiotherapy. This observation is interesting first because of the rare occurrence of Nocardia's peritonitis in the patients on CAPD, and secondly in relation with the species of Nocardia, Nocardia farcinica, less known than Nocardia asteroides but not less pathogenic.

Adult↗

PubMed1976

[Phagocytosis and intracellular proliferation of Nocardia asteroides (strain Weipheld) in cell structures in vitro. 2. Peritoneal macrophages of guinea-pigs (author's transl)].

The study deals with the phagocytosis of Nocardia asteroides (strain Weipheld) and the subsequent intracellular proliferation in peritoneal macrophage cells. Normal, two-stage-immunized and long-term cortison-treated guinea-pig (28 mg cortison / kg weight / day during 30 days) macrophages were used. Further, the cytotoxic effect of Nocardia upon the cells in the peritoneal washing liquid in vitro and the influence of the normal, immune and antimacrophage serum upon the phagocytosis and the intracellular proliferation were studied. Among the cells obtained from the peritoneal washing liquid macrophages were most frequently subject to phagocytosis, leukocytes to a lesser degree. The normal macrophages phagocytized in 14.56% (Fig. 1), macrophages of two-stage-immunized guinea-pigs in 18.21% (Fig. 2) and macrophages from cortison treated guinea-pigs in 12.48% of cases. Intracellular observation showed phagocytized germs after 3 min. of exposure. The course of the intracellular proliferation of Nocardia can be seen in Fig. 3. The phagocytosis index increases slowly in all three groups of macrophages; least so in the immunized macrophages (1.30-after 8 hours). The highest values were obtained in the macrophages of cortison treated guinea-pigs (2.02-after 8 hours). Within 8 hours of exposure the filaments of Nocardia grew through the cell membrane of phagocytizing cells (Figs. 4 A, 4 B). Fig. 5 shows the course of the cytopathogenic effect of Nocardia upon the cells. After 1 hr. the number of dead cells increased from 0.30% to 1.9-3.8%; after 4 hrs. it reached 8.15-9.80%; after 8 hrs. 10.1-14.80%. The highest values were observed in cells from cortison treated guinea-pigs (14.80%). After addition of normal serum (time of phagocytosis 60 min.) normal peritoneal macrophages phagocytized in 13.30% of cases; immune serum stimulated phagocytosis (16.21%); antimacrophage serum significantly reduced phagocytosis (4.10%). The phagocytosis index in peritoneal macrophages with normal and immune serum increased, reaching values of 1.20-1.58 with in 8 hours; the differences were statistically not significant (Fig. 6). A significant increase of proliferation was observed in peritoneal macrophages with antimacrophage serum (2.07 in 8 hours). Neither normal nor immune guine-pig serum influenced the cytopathogenic effect of Nocardia on the cells of peritoneal washing liquid (Fig. 7). The antimacrophage serum had a rapid and expressive effect upon the macrophages (78.23% of cells died after 2 hours, 100% after 3 hours of exposure).

Animals↗

PubMed1993

Circulating antibody to Nocardia in the serum of patients with Parkinson's disease.

Nocardia is a genus of aerobic Gram-positive bacteria which forms filamentous cells that fragmented into rod-shaped or coccoid elements or L-forms. According to the literature N. asteroides causes encephalitis with parkinsonian features, and it may be an unknown etiologic agent causing encephalitis with a parkinsonian syndrome. Because there are no reliable immunologic tests routinely used for diagnosing nocardial infection, less severe CNS infections with nocardiae may not be recognized or may be attributed to unknown or incorrect etiologic agents. Recently, it was shown that sublethal doses of N. asteroides causes an L-DOPA-responsive movement disorder with Lewy-like bodies in mice. In this study, we detected antibodies to nocardia in the serum of patients with Parkinson's disease (PD). Sera was diluted in PBS and added in two-fold dilutions to coccoid and rod-shaped cells of nocardia attached to nonfluorescence slides. IF test demonstrated antibodies to coccoid and rod-shaped cells of nocardia in the serum from 20/20 patients with PD at a titer greater than 1:10, and 14 controls showed 10 positively. The results suggested that not only PD patients but also age-matched healthy volunteers are routinely exposed to and naturally infected with nocardia-related microorganisms. A further reliable immunologic test will be required.

Antibodies, Bacterial↗

PubMed1989

The efficiency of 1 M NH4Cl and 2 M NaCl for the isolation of pathogenic Nocardia from soil.

The present paper reports an improvement to the classical method of the paraffin bait, by the usage of 1 M NH4Cl or 2 M NaCl to eliminate contaminant microflora of soil. The purpose is to introduce a change in the paraffin bait method in order to reduce time required to isolate pathogenic strains of Nocardia from their natural sources. For this study three main criteria were used: a) Determination of the inhibitory effect of different concentrations of salts on soil microflora; b) The tolerance of Nocardia brasiliensis, Nocardia asteroides and Nocardia caviae (Nocardia otitidis caviarum) strains to these chemical inhibitors; c) Determination of the efficiency of salts in the isolation of Nocardia from soil when strains are grown on paraffin baits.

Ammonium Chloride↗

PubMed1996

Occurrence and specificity of human natural and in vitro induced antibodies to Nocardia opaca antigens.

Nocardia opaca, a Gram-positive bacterium, is a potent source of immunostimulatory substances. Screening of sera of adult human donors revealed that all sera tested contained antibodies reactive with isolated Nocardia fractions (Nocardia delipidated cell mitogen, NDCM; Nocardia lysozyme digest, NLD; Nocardia water-soluble mitogen, NWSM; and fraction B). The respective values of reciprocal titres for IgM and IgG were in the range of 100 to 12,800, and 10 to 320 for IgA antibody isotypes, when NLD or fraction B were used as antigens in enzyme-linked immunosorbent assay (ELISA) tests. The level of antibodies directed to NDCM, a potent polyclonal B cell activator, was found to be the lowest. In vitro spontaneous as well as NDCM-induced production of antibodies to NDCM by human peripheral blood lymphocytes involved mainly the IgM class. Western-blot analysis demonstrated that antibodies in normal human sera react with nocardial antigens of molecular mass approximately 60, 40, 20 and 15-10 kDa. The same antigens were also recognized by rabbit and mouse hyperimmune sera, also confirming the immundominancy of these nocardial antigens in other species. The presence of anti-nocardia antibodies in human sera and their production by both stimulated and non-stimulated lymphocytes points to the natural sensitization of humans either by ubiquitous no-cardial components or by cross-reactive bacterial or food antigens.

Adult↗

PubMed1999

Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis and plasmid profile of soil and clinical isolates of Nocardia.

The aim of this study was to develop a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay for generic and species-specific differentiation of Nocardia from other morphologically similar bacterial pathogens. To examine the utility of the PCR-RFLP approach in species identification, genomic DNA was prepared from 40 soil isolates, 10 clinical isolates and 8 reference strains of Nocardia. A set of oligonucleotide primers was designed from the consensus sequence of the highly conserved groEL gene that encodes the 65-kDa heat shock protein (hsp 65). The primers selectively amplified 422 bp DNA from the genomic DNA of all Nocardia species and isolates. The digestion of the amplicons with the restriction enzyme MspI produced DNA fragments that could differentiate between different Nocardia species regardless of their origin. Additionally, the RFLP patterns obtained with restriction enzymes MspI and BsaHI resulted in the differentiation of six Nocardia species which were earlier identified by biochemical tests. Apart from soil isolates of N. asteroides, which had shown some degree of genotypic polymorphism with BsaHI, the remaining taxa yielded more consistent results. Our results on the isolation of plasmids indicated that their occurrence is not a consistent feature in Nocardia species. It is neither related to the source of origin (clinical versus saprobic), nor to virulence, anti-microbial resistance or species specificity.

Humans↗

PubMed2002

[Nocardia farcinica: life-threatening chorioiditis under systemic immunosuppression].

BACKGROUND: Immunosuppressed patients occasionally suffer from a multifocal infection with Nocardia. It is important to distinguish Nocardia farcinica from Nocardia asteroides, because of different sensitivity against antibiotics. PATIENT AND METHODS: A 40-year-old patient with polycystic renal disease successfully underwent a kidney transplantation without complications. Immunosuppression consisted of: corticosteroids, azathioprin and ciclosporin A. Ten weeks later he developed acute choroiditis with consecutive retinal detachment and neovascular glaucoma in one eye. In addition, 14 weeks after transplantation CT scans revealed multiple cerebral abscesses. RESULTS: In the course of the disease Nocardia farcinica (N. f.) was identified by cerebral stereotactic biopsy of a cerebral lesion, histological examination of the enucleated globe and sputum culture. Histologically filamentous, eosinophilic organisms were found. Microbiology identified aerobic actinomycetes in cultures and Nocardia farcinica by PCR. Therapeutically the combination of vancomycin, ampicillin, and sulbactam was successful. CONCLUSION: In immunosuppressed patients Nocardia farcinica can become life-threatening. One of the first manifestations may be a choroiditis. Infection of the respiratory tract followed by hematogenous spread is the common way of systemic nocardiosis. Biopsy followed by identification of species by PCR is recommended because of the specific therapeutic strategies associated with each species.

Adult↗

PubMed2003

Evaluation of RIDOM, MicroSeq, and Genbank services in the molecular identification of Nocardia species.

The molecular identification of Nocardia species, when compared to phenotypic identification, has two primary advantages: rapid turn-around time and improved accuracy. The information content in the 5'-end of the 16S ribosomal RNA gene is sufficient for identification of most bacterial species. An evaluation was performed to demonstrate the quality of results provided by two specialized databases (RIDOM and MicroSeq 500 versions 1.1 and 1.4.3, library version 500-0125, respectively) and the more general GenBank database. In addition, these results were compared with phenotypic identifications. Partial 5'-16S rDNA sequences from 64 culture collection strains (DSM, CIP, JCM, and ATCC) were derived, in duplicate, independently in two laboratories. Furthermore, the sequences and the conventional identification results of 91 clinical Nocardia isolates were determined. With the exception of N. soli and N. cummidelens, all Nocardia type strains were distinguishable using 5'-16S rDNA sequencing. Assuming a normal distribution for the pairwise distances of all unique Nocardia sequences and choosing a reporting criterion of > or = 99.12% similarity for a "distinct species", a statistical error probability of 1.0% can be calculated. When the various databases were searched with the clinical isolate sequences RIDOM gave a perfect match in 71.4% of cases whereas MicroSeq yielded a perfect match in only 26.4%. The GenBank service gave a 100% similarity in 59.3% but in 70.4% of these cases the results obtained were not exclusive for a single Nocardia species. Conventional methods gave a correct identification in 59 cases, although most recent taxonomic changes were not taken into account. The RIDOM service (http://www.ridom-rdna.de/) is in the process of making available a comprehensive and high-quality database for bacterial identification purposes and provides excellent results for the majority of Nocardia isolates.

DNA, Bacterial↗

PubMed1998

Nocardia flavorosea sp. nov.

An actinomycete strain, 'Nocardai flavorosea' JCM 3332, was found to have properties consistent with its classification in the genus Nocardia. An almost complete gene sequence of the 16S rDNA of the strain was determined following cloning and sequencing of the amplified gene. The sequence was aligned with those available for nocardiae and phylogenetic trees were inferred using four tree-making algorithms. The organisms consistently formed a distinct clade with the type strain of Nocardia carnea. However, DNA relatedness experiments showed that the strain and N. carnea DSM 43397T belonged to two distinct genomic species. The organism was also distinguished from representative of all of the validly described species of Nocardia using a combination of phenotypic properties. These genotypic and phenotypic data show that the strain merits recognition as a new species of the genus Nocardia. The name proposed for the new species of is Nocardia flavorosea sp. nov. The type strain is JCM 3332T.

Base Sequence↗

PubMed2002

Nocardia pseudovaccinii sp. nov.

Comparative 16S rDNA studies of Nocardia type and reference strains revealed that strain DSM 43406T, identified as Nocardia vaccinii, was wrongly classified. The strain was aerobic, gram-positive and produced scarce, white, branched aerial mycelium and a beige-red substrate mycelium. The reverse side of the colonies was yellow-orange. It showed chemotaxonomic markers that were consistent with its classification in the genus Nocardia. The mycolic acids had chain lengths from 50 to 58 carbon atoms. The 16S rDNA sequence showed the highest similarity to Nocardia nova (97.7%) and N. vaccinii (97.6%), but the strain could be clearly separated from these species and other members of the N. vaccinii cluster by significant differences in biochemical test results and unique fatty acid and mycolic acid patterns. These data led to the conclusion that the isolate represents a novel species within the genus Nocardia, for which the name Nocardia pseudovaccinii sp. nov. is proposed. The type strain is strain AR 368,38366-20T (= DSM 43406T = NRRL B-24154T).

DNA, Bacterial↗

PubMed2004

Nocardia pigrifrangens sp. nov., a novel actinomycete isolated from a contaminated agar plate.

A polyphasic study was undertaken to establish the taxonomic position of an actinomycete strain isolated from a contaminated agar plate. The strain, designated 7031T, had morphological and chemotaxonomic properties typical of the genus Nocardia. An almost-complete 16S rRNA gene sequence determined for the strain was aligned with available sequences for nocardiae, and phylogenetic trees were inferred using three tree-generating algorithms. Strain 7031T clustered with the type strains of Nocardia carnea and Nocardia flavorosea, showing low 16S rRNA gene sequence similarities to these species (97.2 and 97.5 %, respectively). The strain was also distinguished from the closest species by a range of phenotypic properties. It is proposed that the strain be recognized as a novel species of Nocardia, Nocardia pigrifrangens sp. nov., the type strain of which is 7031T (= AS 4.1808T = JCM 11884T).

Bacterial Typing Techniques↗

PubMed2004

Nocardia xishanensis sp. nov., a novel actinomycete isolated from soil.

The taxonomic position of a soil isolate, strain 276(T), was established using a polyphasic approach. The organism showed a range of chemical and morphological properties consistent with its classification in the genus Nocardia. An almost complete 16S rRNA gene sequence determined for the strain was aligned with corresponding sequences of representatives of the genus Nocardia and related taxa using three tree-making algorithms. The organism formed a distinct phyletic line within the evolutionary radiation occupied by the genus Nocardia and was most closely related to the type strain of Nocardia abscessus. However, the two strains shared a low DNA-DNA relatedness value and were readily distinguished using a combination of phenotypic properties. The combined genotypic and phenotypic data show that strain 276(T) should be assigned to the genus Nocardia as a novel species. The name proposed for this new taxon is Nocardia xishanensis sp. nov. The type strain is 276(T) (=CGMCC 4.1860(T)=JCM 12160(T)).

Bacterial Typing Techniques↗

PubMed2005

Nocardia takedensis sp. nov., isolated from moat sediment and scumming activated sludge.

Chemotaxonomic and morphological characterization of two actinomycete strains, MS1-3T and AS4-2, respectively isolated from moat sediment and scumming activated sludge, was carried out. This characterization clearly demonstrated that strains MS1-3T and AS4-2 belong to the genus Nocardia. 16S rRNA gene sequencing studies showed that these isolates are most closely related to Nocardia beijingensis (98.1-98.3 % similarity), Nocardia brasiliensis (97.9-98.0 %) and Nocardia tenerifensis (97.8-97.9 %). However, the results of DNA-DNA hybridizations and physiological and biochemical tests showed that strains MS1-3T and AS4-2 could be differentiated from their closest phylogenetic relatives both genotypically and phenotypically. It is proposed that the two isolates be classified as representatives of a novel species of Nocardia, Nocardia takedensis sp. nov. The type strain is MS1-3T (=NBRC 100417T=DSM 44801T); AS4-2 (=NBRC 100418=DSM 44802) is a reference strain.

Bacterial Typing Techniques↗

PubMed2006

Characterization of Nocardia plasmid pXT107.

Nocardia, Rhodococcus and Streptomyces, all members of the actinomycetes family, are Gram-positive eubacteria with high G+C content and able to form mycelium. We report here a newly identified plasmid pXT107 of Nocardia sp. 107, one of the smallest circular plasmids found in Nocardia. The complete nucleotide sequence of pXT107 consisted of 4335 bp with 65% G+C content, and encoded one replication extragenic palindromic (Rep) and six hypothetical proteins. The Rep, double-strand origin and single-strand origin of pXT107 resembled those of typical rolling-circle-replication plasmids, such as pNI100 of Nocardia, pRE8424 of Rhodococcus and pIJ101 of Streptomyces. The Escherichia coli-Nocardia shuttle plasmid pHAQ22, containing the rep gene of pXT107, is able to propagate in Nocardia but not in Streptomyces.

Base Sequence↗

PubMed1978

Beta-lactamase production and resistance to beta-lactam antibiotics in Nocardia.

Although ampicillin has been suggested as a useful agent for the treatment of nocardiosis in man, little is known regarding the presence of beta-lactamase in Nocardia or its possible role in determining resistance to ampicillin and the other beta-lactam antibiotics. We have evaluated 55 isolates of Nocardia for susceptibility to five beta-lactam antibiotics and for the presence of beta-lactamase. Nocardia were resistant to penicillin G, cloxacillin, and cefazolin, but 27 and 62% were susceptible to 3.1 and 25 mug of ampicillin per ml, respectively. Almost 90% of these ampicillin-susceptible or intermediate strains were also susceptible to carbenicillin. The combination of ampicillin and cloxacillin was synergistic against many ampicillin-resistant strains. Beta-lactamase was detected in 89% of Nocardia isolates when intact cells were used and in six of six strains after cell fractionation. This beta-lactamase was most active against penicillin G and ampicillin, with lesser activity against carbenicillin and cephaloridine. These studies suggest that beta-lactamase may be present in all clinical isolates of Nocardia and that mechanisms of antimicrobial resistance other than or in addition to beta-lactamase are responsible for resistance of Nocardia to ampicillin and carbenicillin.

Ampicillin↗

PubMed1996

Activities of the glycylcyclines N,N-dimethylglycylamido-minocycline and N,N-dimethylglycylamido-6-demethyl-6-deoxytetracycline against Nocardia spp. and tetracycline-resistant isolates of rapidly growing mycobacteria.

Susceptibilities to the new semisynthetic tetracycline (Tet) compounds N,N-dimethylglycylamido-minocycline (DMG-MINO) and N,N-dimethylglycylamido-6-demethyl-6-deoxytetracycline (DMG-DMDOT) were compared with those to doxycycline, minocycline, and Tet for 198 Tet-resistant (Tetr) and 33 Tet-susceptible (Tets) clinical isolates of rapidly growing mycobacteria (RGM) including the Mycobacterium fortuitum group, Mycobacterium abscessus, Mycobacterium chelonae, and Mycobacterium mucogenicum and 68 isolates belonging to six taxa of Nocardia spp. All Tetr RGM were highly susceptible to the glycylcyclines. The MICs at which 50 and 90% of isolates are inhibited were < or = 0.125 and < or = 0.25 microgram/ml, respectively, for DMG-DMDOT and < or = 0.25 and 1 microgram/ml, respectively, for DMG-MINO. The MIC of DMG-DMDOT at which 50% of Tetr strains are inhibited was the same as that for Tets strains for each of the four taxa of RGM. The new agents were less active against Nocardia spp. MICs of DMG-DMDOT were comparable to those of minocycline except for the MICs for Nocardia brasiliensis sensu stricto, the new taxon Nocardia pseudobrasiliensis, and some isolates of Nocardia nova, against which they were four- to eightfold more active. The MICs of DMG-DMDOT were consistently lower than those of DMG-MINO for RGM. This class of drugs offers exciting therapeutic potential for RGM and for selected species of Nocardia.

Anti-Bacterial Agents↗

PubMed1985

Intracellular acid phosphatase content and ability of different macrophage populations to kill Nocardia asteroides.

It has been reported that the activity of lysosomal acid phosphatase decreases inversely with numbers of ingested virulent Nocardia spp. in normal murine peritoneal and alveolar macrophages. These studies suggested that this relationship correlated with the effectiveness of these macrophage populations in killing Nocardia asteroides. Experiments were designed to determine if acid phosphatase activity is affected by infection with N. asteroides in four different macrophage populations isolated from normal and nocardia-immunized mice. Macrophages were also tested simultaneously for their ability to kill N. asteroides. Peritoneal, alveolar, and splenic macrophages and Kupffer cells were infected in vitro with strains of N. asteroides of differing virulence. Uptake and killing assays were performed. Acid phosphatase levels and numbers of intracellular nocardiae were quantitated in the same macrophages, using a computer-assisted cytophotometry system. Acid phosphatase activity decreased inversely with numbers of intracellular nocardiae in macrophages that could not kill or inhibit this pathogen. Acid phosphatase activity was not significantly changed in macrophages that inhibited growth of, but did not kill, N. asteroides, whereas activity was increased or enhanced in macrophages that killed most of the ingested nocardiae. The order of nocardicidal effectiveness (and resistance to enzyme activity reduction with infection) for normal macrophages was splenic greater than peritoneal greater than alveolar greater than Kupffer. In contrast, the order of these two parameters for macrophages isolated from immunized mice was Kupffer greater than peritoneal greater than alveolar greater than splenic. These results demonstrate that lysosomal acid phosphatase activity is an effective marker of the ability of macrophages to inhibit growth of and kill N. asteroides and that macrophages isolated from different anatomical sites differ functionally from each other with respect to nocardicidal and acid phosphatase activities.

Acid Phosphatase↗

PubMed1991

Use of a siderophore detection medium, ethylene glycol degradation, and beta-galactosidase activity in the early presumptive differentiation of Nocardia, Rhodococcus, Streptomyces, and rapidly growing Mycobacterium species.

A simplified scheme for the presumptive early identification of Nocardia, Rhodococcus, rapidly growing Mycobacterium, and Streptomyces species is presented. The Nocardia and Streptomyces spp. and the Mycobacterium spp. were positive. The spp. were positive for siderophore activity, but only 25% of the Rhodococcus spp. were positive. The Rhodococcus and Mycobacterium spp. were negative for beta-galactosidase, while the Nocardia and Streptomyces spp. were positive. The Nocardia and Streptomyces spp. and the Mycobacterium spp. were negative for ethylene glycol degradation, while 75% of the Rhodococcus spp. were positive. In combination, these tests were useful for differentiating Mycobacterium, Rhodococcus, and Nocardia species but did not differentiate Nocardia from Streptomyces species.

Actinomycetales↗